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MB Sample ID: SA214015
Local Sample ID: | 0_EA_3-neg |
Subject ID: | SU002325 |
Subject Type: | Plant |
Subject Species: | Camelina Sativa |
Age Or Age Range: | 10 days after fertilization |
Species Group: | Developing seeds |
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Collection:
Collection ID: | CO002318 |
Collection Summary: | Plant growth and culture conditions: Plants were grown in greenhouses with day/night temperature maintained at 22/20°C, 40-50% relative humidity, and 16h day/8h night photoperiod. Intact siliques during the seed filling growth stage (15 days after fertilization) were excised and placed in sterile media containing a modified Linsmaier and Skoog medium23,24 with Gamborg’s vitamins (Sigma) and 5 mM MES buffer adjusted to pH 5.8. Fifty mM [U-13C6]glucose was used as labeled substrate, and the composition of the remaining carbon and nitrogen sources represented maternal phloem composition to minimize metabolic perturbation and to maintain pseudo in vivo conditions as previously described25. Silique culturing was performed in a 96-well plate with 0.3 mL of medium and a single silique per well, under continuous light (250 µmol m-2 s-1). Tissue was collected and flash frozen immediately after each time point (2, 4, 8, 16 and 32h). Uncultured siliques excised from the maternal plant were used as unlabeled (0h) controls. Frozen tissue was sectioned, on top of dry ice, to excise embryo from the siliques and to separate cotyledons from the embryo axis. Cotyledon samples were extracted and analyzed for lipids in triplicates. |
Collection Protocol Filename: | 13CLipids_CamelinaSeeds_Methods.docx |
Sample Type: | Seeds |
Collection Location: | Donald Danforth Plant Science Center |
Storage Conditions: | -80℃ |